Method for co-purification of equine neutrophil elastase and myeloperoxidase from a limited blood volume.

[en] Neutrophil myeloperoxidase (MPO) and elastase can be released in severe inflammatory diseases and cause tissue injuries. Equine enzymes have already been individually purified from large blood quantities. We describe the isolation of both enzymes from a same limited blood volume. Both MPO and elastase were extracted by crushing PMN isolated by centrifugation on a percoll-gradient from a 460ml blood collection. MPO and elastase were separated by an ionic exchange chromatography phase and further purified by gel filtration chromatography on Superdex 200 and 75, respectively. Enzymes were identified in the collected fractions by specific enzymatic assays. The final purity was verified by electrophoresis. Specific activity was improved to 19.92 and 34.3x for elastase (final yield: 340mug) and MPO (final yield: 130mug), respectively, during the procedure. Results show the possibility of isolating both enzymes from the same blood sample with a sufficient yield and purity for future studies on their implication and interaction during inflammatory diseases.

Research center :

CORD - Centre de l'Oxygène, Recherche et Développement - ULiège

Disciplines :

Veterinary medicine & animal health
Biochemistry, biophysics & molecular biology

Author, co-author :

de la Rebière de Pouyade, Geoffroy ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux

Serteyn, Didier ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux

Deby, Ginette ; Université de Liège - ULiège > Centre de l'oxygène : Recherche et développement (C.O.R.D.)

Franck, Thierry ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux

Language :

English

Title :

Method for co-purification of equine neutrophil elastase and myeloperoxidase from a limited blood volume.

Publication date :

2009

Journal title :

Research in Veterinary Science

ISSN :

0034-5288

eISSN :

1532-2661

Publisher :

British Veterinary Association, London, United Kingdom

Peer reviewed :

Peer Reviewed verified by ORBi

Funders :

FRIA - Fonds pour la Formation à la Recherche dans l'Industrie et dans l'Agriculture [BE]

Available on ORBi :

since 28 May 2009

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